Michelle L. Delco

Characterization of tissue response to impact loads delivered using a hand-held instrument for studying articular cartilage injury

Objective: The objective of this study was to fully characterize the mechanics of an in vivo impactor and correlate the mechanics with superficial cracking of articular surfaces. Design: A spring-loaded impactor was used to apply energy-controlled impacts to the articular surfaces of neonatal bovine cartilage. The simultaneous use of a load cell and displacement sensor provided measurements of stress, stress rate, strain, strain rate, and strain energy density. Application of India ink after impact was used to correlate the mechanical inputs during impact with the resulting severity of tissue damage. Additionally, a signal processing method to deconvolve inertial stresses from impact stresses was developed and validated. Results: Impact models fit the data well (root mean square error average ~0.09) and provided a fully characterized impact. Correlation analysis between mechanical inputs and degree of superficial cracking made visible through India ink application provided significant positive correlations for stress and stress rate with degree of surface cracking (R2 = 0.7398 and R2 = 0.5262, respectively). Ranges of impact parameters were 7 to 21 MPa, 6 to 40 GPa/s, 0.16 to 0.38, 87 to 236 s−1, and 0.3 to 1.1 MJ/m3 for stress, stress rate, strain, strain rate, and strain energy density, respectively. Thresholds for damage for all inputs were determined at 13 MPa, 15 GPa/s, 0.23, 160 s−1, and 0.59 MJ/m3 for this system. Conclusions: This study provided the mechanical basis for use of a portable, sterilizable, and maneuverable impacting device. Use of this device enables controlled impact loads in vitro or in vivo to connect mechanistic studies with long-term monitoring of disease progression.

Post‐traumatic osteoarthritis of the ankle: A distinct clinical entity requiring new research approaches

The diagnosis of ankle osteoarthritis (OA) is increasing as a result of advancements in non‐invasive imaging modalities such as magnetic resonance imaging, improved arthroscopic surgical technology and heightened awareness among clinicians. Unlike OA of the knee, primary or age‐related ankle OA is rare, with the majority of ankle OA classified as post‐traumatic (PTOA). Ankle trauma, more specifically ankle sprain, is the single most common athletic injury, and no effective therapies are available to prevent or slow progression of PTOA. Despite the high incidence of ankle trauma and OA, ankle‐related OA research is sparse, with the majority of clinical and basic studies pertaining to the knee joint. Fundamental differences exist between joints including their structure and molecular composition, response to trauma, susceptibility to OA, clinical manifestations of disease, and response to treatment. Considerable evidence suggests that research findings from knee should not be extrapolated to the ankle, however few ankle‐specific preclinical models of PTOA are currently available. The objective of this article is to review the current state of ankle OA investigation, highlighting important differences between the ankle and knee that may limit the extent to which research findings from knee models are applicable to the ankle joint. Considerations for the development of new ankle‐specific, clinically relevant animal models are discussed.

Mitochondrial dysfunction is an acute response of articular chondrocytes to mechanical injury

Mitochondrial (MT) dysfunction is known to occur in chondrocytes isolated from end‐stage osteoarthritis (OA) patients, but the role of MT dysfunction in the initiation and early pathogenesis of post‐traumatic OA (PTOA) remains unclear. The objective of this study was to investigate chondrocyte MT function immediately following mechanical injury in cartilage, and to determine if the response to injury differed between a weight bearing region (medial femoral condyle; MFC) and a non‐weight bearing region (distal patellofemoral groove; PFG) of the same joint. Cartilage was harvested from the MFC and PFG of 10 neonatal bovids, and subjected to injurious compression at varying magnitudes (5‐17 MPa, 5‐34 GPa/s) using a rapid single‐impact model. Chondrocyte MT respiratory function, MT membrane polarity, chondrocyte viability, and cell membrane damage were assessed in situ. Cartilage impact resulted in MT depolarization and impaired MT respiratory function within 2 h of injury. Cartilage from a non‐weight bearing region of the joint (PFG) was more sensitive to impact‐induced MT dysfunction and chondrocyte death than cartilage from a weight‐bearing surface (MFC). Our findings suggest that MT dysfunction is an acute response of chondrocytes to cartilage injury, and that MT may play a key mechanobiological role in the initiation and early pathogenesis of PTOA. Clinical significance: Direct therapeutic targeting of MT function in the early post‐injury time frame may provide a strategy to block perpetuation of tissue damage and prevent the development of PTOA.

Sub-critical impact inhibits the lubricating mechanisms of articular cartilage

Although post-traumatic osteoarthritis accounts for a significant proportion of all osteoarthritis, the understanding of both biological and mechanical phenomena that lead to cartilage degeneration in the years to decades after trauma is still lacking. In this study, we evaluate how cartilage lubrication is altered after a sub-critical impact (i.e., an impact to the cartilage surface that produces surface cracking but not full thickness fissuring). Through utilizing a Stribeck-like framework, the elastoviscous transition, we evaluated changes to both the innate boundary lubricating ability of cartilage after impact and also the effectiveness of high viscosity lubricants to lower friction after impact. Increases in boundary friction coincided with changes in lubricin localization after impact. However, larger increases in friction coefficient were observed in mixed-mode lubrication which can be predicted by increases in surface roughness due to cartilage fissuring. The data here reveal distinct mechanisms of cartilage lubrication that can fail after traumatic impact and may explain a key mechanical phenomenon that predisposes cartilage to development of osteoarthritis after injury.

Mitoprotective therapy preserves chondrocyte viability and prevents cartilage degeneration in an ex vivo model of posttraumatic osteoarthritis: MITOPROTECTION IN CARTILAGE

No disease‐modifying osteoarthritis (OA) drugs are available to prevent posttraumatic osteoarthritis (PTOA). Mitochondria (MT) mediate the pathogenesis of many degenerative diseases, and recent evidence indicates that MT dysfunction is a peracute (within minutes to hours) response of cartilage to mechanical injury. The goal of this study was to investigate cardiolipin‐targeted mitoprotection as a new strategy to prevent chondrocyte death and cartilage degeneration after injury. Cartilage was harvested from bovine knee joints and subjected to a single, rapid impact injury (24.0 ±1.4 MPa, 53.8 ± 5.3 GPa/s). Explants were then treated with a mitoprotective peptide, SS‐31 (1µM), immediately post‐impact, or at 1, 6, or 12 h after injury, and then cultured for up to 7 days. Chondrocyte viability and apoptosis were quantified in situ using confocal microscopy. Cell membrane damage (lactate dehydrogenase activity) and cartilage matrix degradation (glycosaminoglycan loss) were quantified in cartilage‐conditioned media. SS‐31 treatment at all time points after impact resulted in chondrocyte viability similar to that of un‐injured controls. This effect was sustained for up to a week in culture. Further, SS‐31 prevented impact‐induced chondrocyte apoptosis, cell membrane damage, and cartilage matrix degeneration. Clinical Significance: This study is the first investigation of cardiolipin‐targeted mitoprotective therapy in cartilage. These results suggest that even when treatment is delayed by up to 12 h after injury, mitoprotection may be a useful strategy in the prevention of PTOA.

Microscale frictional strains determine chondrocyte fate in loaded cartilage

Mounting evidence suggests that altered lubricant levels within synovial fluid have acute biological consequences on chondrocyte homeostasis. While these responses have been connected to increased friction, the mechanisms behind this response remain unknown. Here, we combine a frictional bioreactor with confocal elastography and image-based cellular assays to establish the link between cartilage friction, microscale shear strain, and acute, adverse cellular responses. Our incorporation of cell-scale strain measurements reveals that elevated friction generates high shear strains localized near the tissue surface, and that these elevated strains are closely associated with mitochondrial dysfunction, apoptosis, and cell death. Collectively, our data establish two pathways by which chondrocytes negatively respond to friction: an immediate necrotic response and a longer term pathway involving mitochondrial dysfunction and apoptosis. Specifically, in the surface region, where shear strains can exceed 0.07, cells are predisposed to acute death; however, below this surface region, cells exhibit a pathway consistent with apoptosis in a manner predicted by local shear strains. These data reveal a mechanism through which cellular damage in cartilage arises from compromised lubrication and show that in addition to boundary lubricants, there are opportunities upstream of apoptosis to preserve chondrocyte health in arthritis therapy.